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Selected Works of David M. Ojcius
Presentation
ATP and adenosine receptors modulate IL-1β secretion during Porphyromonas gingivalis infection
Excellence Day
  • Cassio Luiz Coutinho Almeida-da-Silva, University of the Pacific
  • Erivan Schnaider Ramos-Junior, University of the Pacific
  • Ana Carolina Morandini, University of the Pacific
  • Brandon Triell Huynh, University of the Pacific
  • Angela Hwang, University of the Pacific
  • Ozge Carkan, University of the Pacific
  • Shijun Li, University of the Pacific
  • Ryan Park, University of the Pacific
  • David M. Ojcius, University of the Pacific
  • Robson Coutinho-Silva, Universidade Federal do Rio de Janeiro
Lead Author Program & Year
DDS Year 1
Introduction/Context/Diagnosis

Porphyromonas gingivalis (P. gingivalis) is a Gram-negative bacterium strongly associated with periodontitis. This pathogen induces pro- interleukin (IL)-1β synthesis by infected cells, but requires ligation of the P2X7 receptor by extracellular ATP (eATP) to trigger IL-1β release through NLRP3 inflammasome. We already demonstrated that eATP-induced IL- 1β secretion is impaired by P. gingivalis fimbriae via P2X7 receptor in macrophages. Here, we examined the effect of the activation of the purinergic signaling modulating IL-1β production during infection in macrophages.

Methods/Treatment Plan

Bone-marrow derived macrophages were obtained from C57BL/6 or P2X7-/- mice and infected with P. gingivalis or P. gingivalis-nucleoside-diphosphate-kinase (NDK) deficient strain in an MOI of 100 for 6h, 18h and 24h. IL-1β production and secretion was detected by western blotting and ELISA. Antibiotic protection assay quantified bacterial invasion in macrophages. ATP secretion was measured using a bioluminescent reaction. Macrophage ectoenzymes activity was measured by pyrophosphate degradation using a colorimetric assay. Real-time PCR after 2h of infection was performed to analyze adenosine receptors expression.

Results/Outcome

We demonstrated P. gingivalis infection in macrophages requires eATP treatment and signaling via P2X7 receptor to promote IL-1β in 6h, 18h or 24h of infection. We showed P. gingivalis is able to induce pro-IL-1β production independently of P2X7 receptor in macrophages and that the pathogen infect wild-type and P2X7-/- macrophages in similar levels. Next, we showed P. gingivalis induces ATP secretion from macrophages after infection and that P. gingivalis-NDK as long as macrophage ectoenzymes degrade eATP thus favoring production of extracellular adenosine. Adenosine potentiated eATP-induced IL-1β release in infected macrophages via adenosine receptors, probably via A2A receptor.

Significance/Conclusions

Together, our data demonstrate that purinergic signaling via P2X7 and adenosine receptors modulates eATP-induced IL-1β secretion in P. gingivalis-infected macrophages.

Comments/Acknowledgements

This work was supported by FAPERJ scholarship to CLCAS, CNPq, and Funds from the Arthur Dugoni School of Dentistry.

Location
University of the Pacific, Dugoni Dental School, San Francisco, CA
Format
Poster
Poster Session
Faculty, Student, and Staff Presentations
Citation Information
Cassio Luiz Coutinho Almeida-da-Silva, Erivan Schnaider Ramos-Junior, Ana Carolina Morandini, Brandon Triell Huynh, et al.. "ATP and adenosine receptors modulate IL-1β secretion during Porphyromonas gingivalis infection" (2017)
Available at: http://works.bepress.com/david-ojcius/244/